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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Citrate Plasma (50 uL)
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Sensitivity
5 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human Eotaxin
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC21, Quantikine Immunoassay Control Group 4 -
Product Summary
The Quantikine Human Eotaxin Immunoassay is a 3.5 hour solid phase ELISA designed to measure human Eotaxin in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Eotaxin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human Eotaxin accurately. Results obtained measuring natural human eotaxin showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Eotaxin.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 66.8 | 184 | 390 | 67.5 | 193 | 396 |
| Standard Deviation | 2 | 8.4 | 23.5 | 7.8 | 18.2 | 40.2 |
| CV% | 3 | 4.6 | 6 | 11.6 | 9.4 | 10.2 |
Serum, EDTA Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 64.5 | 109 | 377 | 63.4 | 190 | 400 |
| Standard Deviation | 2.5 | 5.8 | 12.8 | 7.3 | 19.6 | 33.5 |
| CV% | 3.9 | 5.3 | 3.4 | 11.5 | 10.3 | 8.4 |
Recovery
The recovery of Eotaxin spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 100 | 86-111 |
| Citrate Plasma (n=5) | 99 | 81-117 |
| EDTA Plasma (n=5) | 99 | 82-115 |
| Serum (n=5) | 95 | 83-109 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of Eotaxin in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: CCL11/Eotaxin
CCL11/Eotaxin is a chemokine that interacts with CCR3 to induce eosinophil, basophil, dendritic cell, and Th2 cell chemoattraction during inflammatory and allergic reactions. Its production is upregulated in airway epithelium, alveolar macrophages, eosinophils, smooth muscle cells, chondrocytes, and fibroblasts following allergen or inflammatory challenge. CCL11 is also involved in the growth of myeloid cell progenitors and the differentiation of mast cells.
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